Biochemical, proteomic and microscopic insights into regulation of nucleolar structure and function

对核仁结构和功能调节的生物化学、蛋白质组学和微观见解

• 批准号: RGPIN-2015-06674
• 负责人: TrinkleMulcahy, Laura
• 金额: $ 2.48万
• 依托单位: University of Ottawa
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2016
• 资助国家: 加拿大
• 起止时间: 2016-01-01 至 2017-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=614291
• 关键词: Biochemical; proteomic; microscopic; insights; into

The mammalian nucleolus is a non membrane-bound nuclear organelle that forms around tandem repeats of ribosomal DNA and is best known for its essential role in the coordination of ribosome biogenesis, which supplies the cell with the ribosome complexes required for protein synthesis. This structure is dynamic, disassembling and reassembling throughout each cell cycle in mammalian cells and reorganizing its internal structure in response to various cellular stresses. Recent biochemical and proteomic analyses have revealed a much wider nucleolar functional complexity than was previously appreciated, linking this organelle to roles in cell cycle regulation, DNA damage repair, telomere metabolism, RNA processing and coordination of the cellular response to various stresses. Given that nucleolar dysfunction has been linked to several disease states, including cancer, accelerated aging and viral infection, it is clear that precise coordination of these diverse individual pathways is essential for its proper function.     Despite major advances in recent years, including a more detailed catalogue of nucleolar protein, DNA and RNA components, important questions remain to be answered with regard to which specific factors provide the nucleolus with its structural integrity and how its dynamic organization is regulated. Much also remains to be learned about the full range of biological processes that occur within, or involve, the nucleolus, and the relationship between sub-nucleolar structure and specific functions at the molecular level.     Our interest lies primarily in the structure/function relationship of non-canonical nucleolar components (i.e. those not involved directly in ribosome biogenesis), and we hypothesize that their functions fit within a defined architecture, similar to the structure/function relationship observed for components involved in ribosome biogenesis pathways. The main objectives of this project are thus to characterize nucleolar proteins that mediate non-ribosomal functions and define the regulatory mechanisms that underlie this organelle’s coordinated organization, and we propose to apply a combination of powerful new super-resolution imaging and quantitative proteomic technologies to carry them out.     Understanding the regulation of nucleolar assembly/maintenance and function remains a fundamental question in cell biology. Given that a detailed description of the protein content of the nucleolus has emerged over the past few years and yet we still do not know how their diverse functions are regulated (and in some cases, what their functions actually are) this structure clearly remains a source of significant new discoveries and undoubtedly some more surprises for the foreseeable future.

哺乳动物的核OLUS是一种非膜结合的核细胞器，周围形成核糖体DNA的串联重复序列，以其在核糖体生物发生配位中的重要作用而闻名，该核糖体生物发生在核糖体生物发生过程中，为细胞提供蛋白质合成所需的核糖体复合物。在哺乳动物细胞中，整个细胞周期中，这种结构是动态的，分解和重新组装的，并对各种细胞应激进行了重组。 最近的生化和蛋白质组学分析表明，核功能的复杂性比以前所欣赏的核能复杂性更大，将这种细胞器与细胞周期调节，DNA损伤修复，端粒代谢，RNA代谢，RNA处理以及细胞反应与各种应力的配位的作用联系起来。鉴于核功能障碍已与几种疾病状态有关，包括癌症，加速衰老和病毒感染，很明显，这些潜水员个体途径的精确协调对于其适当的功能至关重要。 尽管近年来取得了重大进展，包括更详细的核蛋白，DNA和RNA成分目录，但重要的问题仍有待回答，这些问题还需要解决哪些特定因素为其结构完整性以及如何调节其动态组织。关于在分子水平上发生或涉及的核olus以及亚核结构与特定功能之间的关系的全部生物学过程以及涉及的全部生物过程。 我们的兴趣主要是对非规范核成分的结构/功能关系（即，那些不直接参与核糖体生物发生），我们假设它们的功能适合定义的结构，类似于与核糖体生物发生途径有关的组件观察到的结构/功能关系。因此，该项目的主要目标是表征核蛋白，这些核蛋白质介导非核心体功能并定义了该细胞器协调组织的基础的调节机制，我们建议将强大的新型超分辨率成像和定量蛋白质组学技术的组合结合起来。 了解核装配/维护和功能的调节仍然是细胞生物学的基本问题。鉴于过去几年中出现了对核olus蛋白质含量的详细描述，但我们仍然不知道如何调节其潜水功能（在某些情况下，实际上是什么功能），这种结构显然仍然是重大新发现的来源，无疑是可预见的未来的更多惊喜。